Chemiluminescence immunodetection kit for detecting cyproheptadine

A chemiluminescence immunoassay and detection kit technology, applied in the field of immunology detection, can solve the problems of low false positive rate of accurate detection, biased experimental results, and complicated operation steps, so as to ensure long-term effectiveness, simple instrument, and increase sensitivity. Effect

Inactive Publication Date: 2016-02-03
JIANGSU WISE SCI & TECH DEV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

High-performance liquid chromatography (HPLC) has the characteristics of high detection accuracy and low false positive rate; the main determination of high-performance liquid chromatography (HPLC) is that the instrument is expensive, the operation is cumbersome, time-consuming, and the detection cost is high
Gas chromatography-mass spectrometry (GC-MS) method has high sensitivity and low false positive rate; however, the sample of this method needs to be derivatized, which will cause deviations in the experimental results
Liquid chromatography-mass spectrometry (LC-MS) can detect urine, blood, hair and other samples, but similar to the above method, the experimental operation steps are cumbersome and the detection cost is high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] 1. Preparation of each component of the kit

[0040] (1) Preparation of cyproheptadine hapten: acidify cyproheptadine, react with sodium nitrite in a low-temperature environment without light at 4°C to generate an intermediate containing diazonium cations. The diazotized cyproheptadine is used as a hapten for subsequent synthesis of immune antigens and coating antigens;

[0041] (2) Preparation of cyproheptadine-bovine serum albumin (BSA) immunogen: coupling cyproheptadine and bovine serum albumin (BSA) by diazotization method to obtain immune antigen;

[0042] Preparation of cyproheptadine-oval serum albumin (OVA) coated antigen: Coupling cyproheptadine and egg serum albumin (OVA) by diazotization method to obtain coated antigen;

[0043]Preparation of cyproheptadine-peroxidase-labeled antibody: For 6-8 week-old female BALB / c mice (body weight 18-20g), the high-dose immunization scheme is as follows: 160 μg cyproheptadine-BSA and other Mix Freund's complete adjuvant ...

Embodiment 2

[0078] The chemiluminescent immunoassay kit for detecting cyproheptadine includes the following components:

[0079] (1) 96-well opaque white ELISA plate (8 wells x 12 strips) coated with cyproheptadine-mouse serum albumin conjugate, vacuum-sealed in an aluminum foil bag;

[0080] (2) 6 bottles of cyproheptadine standard solution, the concentrations are respectively:

[0081] 0ng / mL, 0.02ng / mL, 0.1ng / mL, 0.5ng / mL, 2.5ng / mL, 12.5ng / mL

[0082] (3) cyproheptadine-horseradish peroxidase labeled antibody solution;

[0083] (4) Luminescence substrate A solution (luminol and enhancer), luminescence substrate B solution (urea hydrogen peroxide);

[0084] (5) 2 times concentrated sample diluent. Before use, please dilute 1:1 (1 part of concentrated diluent + 1 part of deionized water) to become the working sample diluent, and the diluted working sample diluent is 0.05mol / L, pH7.4 Tris-HCl buffer ;

[0085] (6) 20 times concentrated washing solution. Before use, please dilute at ...

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Abstract

The invention provides a chemiluminescence immunodetection kit for detecting cyproheptadine, and belongs to the field of immunological detection. The kit consists of a non-transparent white elisa plate coated with a cyproheptadine-carrier protein conjugate, a cyproheptadine standard product, a cyproheptadine-peroxidase labeled antibody working solution, a luminous substrate solution, a concentrated sample diluting solution and a concentrated cleaning solution. The cyproheptadine-carrier protein conjugate is prepared from cyproheptadine and carrier protein through a mixed anhydride method or a carbodiimide method, and the concentrated cleaning solution comprises 0.05% Tween-20. Compared with the conventional enzyme linked immunosorbent assay method, the kit provided by the invention is higher in sensitivity, short in detection time and low in cost, and can be used for detecting the yproheptadine residual amount of samples of animal urine, blood samples, tissue, internal organs and the like.

Description

technical field [0001] The invention relates to a chemiluminescent immunoassay kit for detecting cyproheptadine, which is used for detecting the content or residual amount of cyproheptadine in animal tissues, viscera, blood, urine, feed, and feed raw materials. It belongs to the field of immunological detection. Background technique [0002] Cyproheptadine (CYP) is a kind of human drug, which is clinically used to treat diseases such as dermatitis caused by allergic reactions. Because the drug can inhibit the satiety center of the hypothalamus and stimulate appetite, it can promote animal growth. . In 2010, Shanghai Veterinary Medicine and Feed Testing Institute discovered for the first time that CYP was illegally used by some illegal animal husbandry enterprises as a new type of breeding accelerator, and the residue of CYP in animal products was extremely harmful to the health of consumers. At the end of 2010, China's Ministry of Agriculture announced No. 1519 "Substances...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/577
Inventor 洪霞薛永来
Owner JIANGSU WISE SCI & TECH DEV
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